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WO2006005593B1 - Method for the simple and rapid detection of cells and biomolecules by means of paramagnetic particles - Google Patents

Method for the simple and rapid detection of cells and biomolecules by means of paramagnetic particles

Info

Publication number
WO2006005593B1
WO2006005593B1 PCT/EP2005/007571 EP2005007571W WO2006005593B1 WO 2006005593 B1 WO2006005593 B1 WO 2006005593B1 EP 2005007571 W EP2005007571 W EP 2005007571W WO 2006005593 B1 WO2006005593 B1 WO 2006005593B1
Authority
WO
WIPO (PCT)
Prior art keywords
microbeads
coated
sample
antibody
detected
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/EP2005/007571
Other languages
German (de)
French (fr)
Other versions
WO2006005593A1 (en
Inventor
Holger Kiesewetter
Abdulgabar Salama
John Adam
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to EP05772506A priority Critical patent/EP1766401B8/en
Priority to US11/632,386 priority patent/US20100009383A1/en
Priority to JP2007520739A priority patent/JP2008506132A/en
Priority to DK05772506T priority patent/DK1766401T3/en
Priority to DE502005006950T priority patent/DE502005006950D1/en
Publication of WO2006005593A1 publication Critical patent/WO2006005593A1/en
Publication of WO2006005593B1 publication Critical patent/WO2006005593B1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
    • G01N33/587—Nanoparticles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/005—Pretreatment specially adapted for magnetic separation
    • B03C1/01—Pretreatment specially adapted for magnetic separation by addition of magnetic adjuvants
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/28—Magnetic plugs and dipsticks
    • B03C1/288—Magnetic plugs and dipsticks disposed at the outer circumference of a recipient
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/30—Combinations with other devices, not otherwise provided for
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C2201/00—Details of magnetic or electrostatic separation
    • B03C2201/18—Magnetic separation whereby the particles are suspended in a liquid
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C2201/00—Details of magnetic or electrostatic separation
    • B03C2201/26—Details of magnetic or electrostatic separation for use in medical or biological applications

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Cell Biology (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nanotechnology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

The invention relates to a simple and rapid method for the detection of cells and biomolecules by means of paramagnetic particles (beads) without separating the detected biomolecules or target cells from the beads before evaluating the specific bond.

Claims

GEÄNDERTE ANSPRÜCHE CHANGED CLAIMS [beim Internationalen Büro am 07. Februar 2006 (07.02.2006) eingegangen; ursprüngliche Ansprüche 1, 3, 5, 10 und 16 geändert; ursprüngliche Ansprüche 2, 8 und 9 gestrichen ; alle weiteren Ansprüche unverändert (2 Seiten)][received at the International Bureau on 7 February 2006 (07.02.2006); original claims 1, 3, 5, 10 and 16 changed; original claims 2, 8 and 9 deleted; all other claims unchanged (2 pages)] +ERKLÄR.UNG+ ERKLÄR.UNG 1. Verfahren zum Nachweis von ausgewählten Zellen (ZZ) oder Biomolekülen in einer Probe, dadurch gekennzeichnet dass man a) paramagnetische Mikrobeads mit einem gegen ein Merkmal der Biomolekü¬ le oder Zellen gerichteten spezifischen Detektionsmolekül beschichtet, b) die Probe mit den beschichteten Mikrobeads in Kontakt bringt, c) die Probe von den nun beladenen Mikrobeads mit Hilfe eines anzulegenden Magneten entfernt, d) die beladenen und von der Probe entfernten Mikrobeads in einem Puffer oder einer Salzlösung resuspendiert, wobei das Resuspensions-Volumen kleiner ist als das ursprüngliche Probenvolumen, e) die in d) erhaltene Mikrobeads-Suspension auf eine Gelkarte schichtet, f) die mit der Mikrobeads-Suspension beladene Gelkarte zentifugiert, und g) anhand des Vorliegens einer Agglutinierung zwischen den Biomolekülen und/oder den ZZ und den beschichteten Mikrobeads direkt das Vorhanden¬ sein des nachzuweisenden Biomoleküls bzw. der nachzuweisenden Zelle vi¬ suell oder photometrisch bestimmt.1. A method for the detection of selected cells (ZZ) or biomolecules in a sample, characterized in that a) paramagnetic microbeads coated with a directed against a characteristic of Biomolekü¬ le or cells specific detection molecule, b) the sample with the coated microbeads in C) removes the sample from the now loaded microbeads with the aid of a magnet to be applied, d) resuspends the loaded and removed microbeads in a buffer or saline solution, the resuspension volume being less than the original sample volume, e ) coating the microbeads suspension obtained in d) on a gel card, f) centrifuging the gel card loaded with the microbeads suspension, and g) directly presenting the presence of agglutination between the biomolecules and / or the ZZ and the coated microbeads be the biomolecule to be detected or the cell to be detected vi¬ suell or p determined by hotometry. 3. Verfahren nach Anspruch 1, dadurch gekennzeichnet, dass die Mikrobeads eine Größe zwischen 0,1 und 5 /xm, bevorzugt 2,5 und 3 /im aufweisen und bevorzugt farbig oder mit Europium markiert sind.3. The method according to claim 1, characterized in that the microbeads have a size between 0.1 and 5 / xm, preferably 2.5 and 3 / im and are preferably colored or marked with europium. 5. Verfahren nach Anspruch 4, dadurch gekennzeichnet, dass die Nanobeads eine Größe von 50 nm bis 0,5 μm, bevorzugt etwa 100 ran aufweisen.5. The method according to claim 4, characterized in that the nanobeads have a size of 50 nm to 0.5 microns, preferably about 100 ran. 10. Verfahren nach einem der vorstehenden Ansprüche, dadurch gekennzeichnet, dass für die Schritte e) bis g) ein Coombs-Röhrchen statt einer Gelkarte verwendet wird. 6. Verfahren zum Nachweis von ausgewählten Zellen (ZZ) oder Biomolekülen in einer Probe, dadurch gekennzeichnet dass man a) paramagnetische Mikrobeads mit einem gegen ein Merkmal der Biomolekü¬ le oder Zellen gerichteten spezifischen Antikörper beschichtet, b) ein Antigen vorlegt, gegen das der nachzuweisende Antikörper gerichtet ist, c) ein Trägermittel mit einem anti-Spezies X- Antikörper beschichtet, wobei Spezies X die Spezies ist, aus der der nachzuweisende Antikörper stammt, und sich diese Spezies von der Spezies unterscheidet, aus der der Antikör¬ per stammt, mit dem die Mikrobeads aus Stufe a) beschichtet sind, d) die beschichteten Mikrobeads aus Stufe a) mit dem Antigen aus Stufe b) in Kontakt bringt, ohne dass ein Kontakt zwischen der Probe und dem Trä¬ germittel mit dem anti-Spezies X- Antikörper möglich ist, e) die Probe mit den beschichteten Mikrobeads in Kontakt bringt, f) die Probe von den nun beladenen Mikrobeads mit Hilfe eines anzulegenden Magneten entfernt, wobei der Magnet so angelegt wird, dass bedingt durch die magnetische Kraft die Konglomerate aus beschichteten Beads, Antigen und nachzuweisendem Antikörper zu dem mit anti-X beschichteten Trägermittel bewegt werden, wobei das Vorhandensein des nachzuweisenden Antikörpers in der Probe durch die Ausbildung einer spezifischen Bindung zwischen diesem nachzuweisenden Antikörper und dem anti-X Antikörper bestimmt wird. 10. The method according to any one of the preceding claims, characterized in that for the steps e) to g) a Coombs tube is used instead of a gel card. 6. A method for the detection of selected cells (ZZ) or biomolecules in a sample, characterized in that a) paramagnetic microbeads coated with a directed against a characteristic of Biomolekü¬ le or cells specific antibodies, b) presents an antigen against which the c) a carrier is coated with an anti-species X antibody, wherein species X is the species from which the antibody to be detected is derived, and this species differs from the species from which the antibody originates; with which the microbeads from stage a) are coated, d) the coated microbeads from stage a) are brought into contact with the antigen from stage b), without any contact between the sample and the carrier medium having the anti-species X- Antibody is possible, e) brings the sample with the coated microbeads in contact, f) remove the sample of the now loaded microbeads using a magnet to be applied wherein the magnet is applied so that due to the magnetic force the conglomerates of coated beads, antigen and antibody to be detected are moved to the anti-X coated carrier, the presence of the antibody to be detected in the sample being due to the formation of a specific antibody Binding between this antibody to be detected and the anti-X antibody is determined.
PCT/EP2005/007571 2004-07-12 2005-07-12 Method for the simple and rapid detection of cells and biomolecules by means of paramagnetic particles Ceased WO2006005593A1 (en)

Priority Applications (5)

Application Number Priority Date Filing Date Title
EP05772506A EP1766401B8 (en) 2004-07-12 2005-07-12 Method for the simple and rapid detection of cells and biomolecules by means of paramagnetic particles
US11/632,386 US20100009383A1 (en) 2004-07-12 2005-07-12 Method for the simple and rapid detection of cells and biomolecules by means of paramagnetic particles
JP2007520739A JP2008506132A (en) 2004-07-12 2005-07-12 Simple and fast method for detecting cells and biomolecules with paramagnetic particles
DK05772506T DK1766401T3 (en) 2004-07-12 2005-07-12 Method for simple and rapid detection of cells and biomolecules by means of paramagnetic particles
DE502005006950T DE502005006950D1 (en) 2004-07-12 2005-07-12 PROCESS FOR THE EASY AND FAST DETECTION OF CELLS AND BIOMOLECULES WITH THE HELP OF PARAMAGNETIC PARTICLES

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102004033811A DE102004033811A1 (en) 2004-07-12 2004-07-12 Method for the simple and rapid detection of cells and biomolecules with the aid of paramagnetic particles
DE102004033811.6 2004-07-12

Publications (2)

Publication Number Publication Date
WO2006005593A1 WO2006005593A1 (en) 2006-01-19
WO2006005593B1 true WO2006005593B1 (en) 2006-03-30

Family

ID=35094256

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2005/007571 Ceased WO2006005593A1 (en) 2004-07-12 2005-07-12 Method for the simple and rapid detection of cells and biomolecules by means of paramagnetic particles

Country Status (9)

Country Link
US (1) US20100009383A1 (en)
EP (1) EP1766401B8 (en)
JP (1) JP2008506132A (en)
AT (1) ATE426810T1 (en)
DE (2) DE102004033811A1 (en)
DK (1) DK1766401T3 (en)
ES (1) ES2321740T3 (en)
PT (1) PT1766401E (en)
WO (1) WO2006005593A1 (en)

Families Citing this family (7)

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US20080097774A1 (en) * 2006-08-29 2008-04-24 Rawls-Meehan Martin B Using a software application to configure a foam spring mattress
ITTO20121085A1 (en) * 2012-12-17 2014-06-18 Davide Maselli DEVICE AND METHOD FOR CELL SELECTION
DE102013106432A1 (en) * 2013-06-20 2014-12-24 Endress + Hauser Conducta Gesellschaft für Mess- und Regeltechnik mbH + Co. KG Optical indicator unit and device and method for determining a physico-chemical property of a process medium in a process plant
KR102323205B1 (en) 2014-08-22 2021-11-08 삼성전자주식회사 Apparatus for separating target matter and Method for separating target matter
US11293055B2 (en) 2015-07-13 2022-04-05 National Cancer Center Nucleic acid detection kit and nucleic acid detection method using nanoparticles
KR101817155B1 (en) * 2015-07-13 2018-01-11 국립암센터 Kit and Method for detecting nucleic acids using nanoparticles
EP3969176A4 (en) 2019-05-14 2023-06-07 The Regents of the University of California PLATFORM FOR THE DETERMINISTIC ARRANGEMENT OF MICROFLUIDIC DROPLETS

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Also Published As

Publication number Publication date
EP1766401A1 (en) 2007-03-28
US20100009383A1 (en) 2010-01-14
WO2006005593A1 (en) 2006-01-19
DK1766401T3 (en) 2009-06-29
EP1766401B1 (en) 2009-03-25
PT1766401E (en) 2009-06-08
EP1766401B8 (en) 2009-07-08
ES2321740T3 (en) 2009-06-10
ATE426810T1 (en) 2009-04-15
DE502005006950D1 (en) 2009-05-07
DE102004033811A1 (en) 2006-02-02
JP2008506132A (en) 2008-02-28

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